This invention is in the field of developmental biology and cell biology. Specifically, this invention relates to a population of Mxc3xcllerian duct-derived epithelial cells that are capable of differentiating into uterine, oviductal, vaginal, and cervical cells, methods of isolating the Mxc3xcllerian duct-derived epithelial cells, characterization of Mxc3xcllerian duct-derived epithelial cells, and uses of the Mxc3xcllerian duct-derived epithelial cells.
In the past few decades, a substantial amount of time and effort has been put into researching the development of female reproductive organs. The impetus behind the research varies from laboratory to laboratory, however, all the research efforts address important common issues relating to the overall health of women. Some of these issues include: cervical cancer, infertility, endometriosis, uterine cancer, ectopic pregnancies, ovarian cysts, and uterine fibroids. Cervical cancer, for example, is a particularly important topic for women""s health considering that cervical cancer is the second most common cancer among women worldwide with approximately 450,000 new cases being diagnosed annually and that almost 200,000 deaths are due to cervical cancer. Pisani et al. Int. J. Cancer 55: 891-903 (1993). Although the etiological cause of cervical cancer remains unknown today, there are many reports that infection with human papillomavirus, in particular, HPV-16 and HPV-18, may be the cause for the development of cervical cancer. Although cervical cancer research has accomplished progress in the past, some of the most critical work is impeded by a lack of human tissue models. Likewise, research relating to ovarian cancers, uterine cancer, uterine fibroids, or endometriosis would benefit greatly from having human tissue models of the cervix, uterus, oviduct (fallopian tube), and vagina.
The cervix, uterus, oviduct, and part of the vagina of the female reproductive system are formed early in embryogenesis from Mxc3xcllerian ducts, also known as paramesonephric ducts. In human embryos, a primordial gonadal ridge develops into a primitive gonad. At about the seventh week of gestation, both sexes have primordial genital ducts and a primitive gonad which develops into a cortex and a medulla. In genetic females, the cortex develops into ovaries and the medulla regresses. In contrast, the medulla develops into testes and the cortex regresses in genetic males. As development of a human embryo progresses, Mxc3xcllerian ducts in males begin to regress with the secretion of Mxc3xcllerian inhibiting substance (or MIS). Ganong, William F. Review of Medical Physiology, Chapter 23 xe2x80x9cThe Gonads: Development and Function of the Reproductive Systemxe2x80x9d, Fifteenth Edition, Appleton and Lange (1991). The Mxc3xcllerian duct is either of the two paired embryonic tubes extending along the mesonephros roughly parallel to the mesonephric duct and emptying into the cloaca. In females, the upper parts of the Mxc3xcllerian duct form the oviducts, while the lower parts fuse to form the uterus, cervix, and part of the vagina.
Previous work on Mxc3xcllerian ducts have focused on anatomical and structural characteristics of Mxc3xcllerian ducts. For example, one study revealed that the movements of Mxc3xcllerian ridges and the immunohistochemical staining of Mxc3xcllerian ducts in avians closely resemble that seen in human. Jacob M., et. al. Cells Tissues Organs 164(2), 63-81, (1999). In another study, human fetuses were examined by ultrasound to study the developing urogenital tracts. Lawrence W. D., et. al. American Journal of Obstetrics and Gynecology 167(1), 185-193, (1992). Other studies have focused on gene expression patterns in the developing fetus. Pellegrini M. et. al. Anat. Embryol. 196(6). 427-433, (1997). While important in their respective scopes, these studies do not provide any teachings for methods of isolating Mxc3xcllerian duct-derived epithelial cells, nor do they provide any teaching for methods for culturing Mxc3xcllerian duct-derived epithelial cells such that the cells retain their pluripotent potential. There are very few, if any, reports of Mxc3xcllerian duct-derived epithelial cells that have been isolated and even fewer reports of Mxc3xcllerian duct-derived epithelial cells that have pluripotent potential to differentiate into uterine, cervical, oviductal, and vaginal cells. Accordingly, there exists a need for the discovery of a population of Mxc3xcllerian duct-derived epithelial cells as well as methods of isolating and characterizing Mxc3xcllerian duct-derived epithelial cells. The invention disclosed herein fulfills these needs and discloses additional methods of use as well.
In one aspect, the invention relates to a population of substantially pure human Mxc3xcllerian duct-derived epithelial cells that have a pluripotent capability to differentiate into oviductal, uterine, vaginal, or cervical cells.
In another aspect, the invention relates to methods of isolating a population of substantially pure human Mxc3xcllerian duct-derived epithelial cells that have the pluripotent capability to differentiate into oviductal, uterine, vaginal, and cervical cells.
In yet another aspect, the invention relates to methods of maintaining a population of substantially pure human Mxc3xcllerian duct-derived epithelial cells that have the pluripotent capability to differentiate into oviductal, uterine, vaginal, and cervical cells and maintaining or culturing these Mxc3xcllerian duct-derived epithelial cells under culture conditions sufficient to allow the Mxc3xcllerian duct-derived epithelial cells to retain their pluripotent capacity.
In still another aspect, the invention relates to methods of providing a source of immunogen to a heterologous recipient and the uses of a substantially pure population of Mxc3xcllerian duct-derived epithelial cells as an immunogen.
In still another aspect of this invention, the invention relates to methods of generating a human tissue model of Mxc3xcllerian duct-derived cells or cells differentiated from Mxc3xcllerian duct-derived cells (i.e. oviductal, uterine, vaginal, and cervical cells) using a substantially pure population of Mxc3xcllerian duct-derived epithelial cells or cell differentiated therefrom as a source of Mxc3xcllerian duct-derived cells and administering the Mxc3xcllerian duct-derived epithelial cells into a non-human, mammalian recipient.
In another aspect of this invention, the invention relates to methods of providing cell therapy whereby a substantially pure population of human Mxc3xcllerian duct-derived epithelial cells or cell differentiated therefrom are introduced into a recipient.
In another aspect of this invention, the invention relates to methods of providing a means for developing pharmaceutical drugs wherein a substantially pure population of human Mxc3xcllerian duct-derived epithelial cells is used as a source of Mxc3xcllerian duct-derived biological components in which one or more of these Mxc3xcllerian duct-derived biological components are the targets of the drugs that are being developed.
In another aspect of this invention, the invention relates to methods of providing bioassay development wherein a substantially pure population of human Mxc3xcllerian duct-derived epithelial cells are used as a source of nucleic acids or proteins and wherein these nucleic acids or proteins are used as one or more principal components in a bioassay or the development of a bioassay.